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目的:探讨3-磷酸甘油醛脱氢酶(glyceraldehyde-3-phophoate de-hydrogenase,GAPDH)是否参与肿瘤坏死因子相关凋亡诱导配体(tumornecrosis factor related apoptosis indu-cing ligand,TRAIL)诱导的甲状腺癌细胞凋亡。方法:光学显微镜、流式细胞仪、实时荧光定量RT-PCR、蛋白质印迹法、台盼蓝染色和共聚焦显微镜分别用于检测细胞生存状态、凋亡、GAPDH mRNA与蛋白表达、细胞活性和核内GAPDH免疫染色表达。结果:对照组和2 ng/mL TRAIL处理组细胞生长状态良好,而其他处理组较差。TRAIL诱导凋亡和GAPDHmRNA均呈剂量依赖性。20 ng/mLTRAIL处理的FRO细胞中,GAPDHmRNA显著增加始于2 h,8 h达平台;GAPDH蛋白核转移始于2 h,8和12 h更为显著;2 h时无细胞死亡;GAPDH核染阳性细胞显著增加始于4 h,12 h时可占大部分。结论:GAP-DH可能介入了TRAIL诱导的细胞凋亡。
Objective: To investigate whether glyceraldehyde-3-phophoate de-hydrogenase (GAPDH) is involved in thyroid cancer induced by tumor necrosis factor related apoptosis inducing ligand (TRAIL) Apoptosis. Methods: The cell viability, apoptosis, GAPDH mRNA and protein expression, cell viability and nucleus were detected by light microscopy, flow cytometry, real-time RT-PCR, Western blotting, trypan blue staining and confocal microscopy. Within GAPDH immunostaining. Results: The cells in the control group and 2 ng / mL TRAIL group grew well, while the other treatment groups were poor. TRAIL induced apoptosis and GAPDH mRNA in a dose-dependent manner. In 20 ng / mLTRAIL-treated FRO cells, GAPDHmRNA increased significantly from 2 h to 8 h, while GAPDH protein nuclear transfection began at 2 h, more significantly at 8 and 12 h, and no cell death at 2 h. Positive cells increased significantly from 4 h, 12 h can account for the majority. Conclusion: GAP-DH may be involved in TRAIL-induced apoptosis.